应用基因芯片筛选转染FOXO1后A549细胞差异表达基因  

Screening of Genes Differentially Expressed in A549 Cells Transfected with FOXO1 Using DNA Microarray

作　　者：孙青峰[1,2] 李理[2] 张安兵[2] 黄文杰[1,2] 

机构地区：[1]广东药科大学研究生学院,广东广州510006 [2]广州军区广州总医院呼吸内科,广东广州510010

出　　处：《中国呼吸与危重监护杂志》2016年第4期368-373,共6页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：国家自然科学基金面上项目(编号:81070003);国家自然科学基金青年基金(编号:81200002)

摘　　要：目的研究转染叉头蛋白O1(FOXO1)后A549细胞基因表达谱的改变,为深入探讨FOXO1在急性肺损伤中的作用机制提供线索。方法应用肿瘤坏死因子α(TNF-α)刺激A549细胞,应用脂质体介导的转染方法将真核表达载体GV230-FOXO1转染入经TNF-α诱导的A549细胞,再提取细胞的总RNA,应用基因芯片技术进行分析。结果成功构建并验证了FOXO1真核表达载体,提取了高质量的mRNA进行芯片分析,并通过RNA质量控制(RNA QC)验证。芯片分析结果显示差异表达基因中上调的基因有317个,下调的基因有237个。这些差异表达的基因功能涉及细胞增殖、凋亡、分化等多个方面。结论应用基因芯片技术可成功筛选转染FOXO1基因后A549细胞的差异表达基因。FOXO1在急性肺损伤中可能通过改变细胞的增殖、凋亡与分化等水平来影响疾病的进程。Objective To study the differences in gene expression in A549 cells transfected with Forkhead box protein O1（ FOXO1）,and provide clues to further exploring the mechanism of FOXO1 in acute lung injury. Methods After using TNF-α to stimulate A549 cells,the eukaryotic expression vector GV230-FOXO1 was transfected into A549 cells by using lipofectamine transfection reagent. The RNA was isolated and differentially expressed genes were screened with high-throughout DNA microarray. Results The eukaryotic expression vector GV230-FOXO1 was successfully constructed and verified. High quality mRNA was isolated and prepared for microarray screening,which passed RNA quality control. The DNA microarray data indicated that 317 genes were up-regulated and 237 genes were down-regulated in A549 cells transfected with FOXO1. The function of these differentially expressed genes involved in many aspects,such as proliferation,apoptosis and differentiation. Conclusions Differentially expressed genes in A549 cells transfected with FOXO1 can be successfully screened by using DNA microarray. FOXO1 may influence the progression of the disease by changing the level of cell proliferation,apoptosis and differentiation in acute lung injury.

关 键 词：叉头蛋白O1 基因芯片 A549细胞 急性肺损伤 

分 类 号：R563[医药卫生—呼吸系统]